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Biochemical  Studies  of  Beryllium 
Sulfate. 


DISSERTATION 


SUBMITTED   IN   PARTIAL   FULFILMENT   OF  THE    REQUIRE- 
MENTS  FOR  THE  DEGREE  OF    DOCTOR   OF   PHIL- 
OSOPHY IN  THE  FACULTY  OF  PURE  SCIENCE 
OF   COLUMBIA   UNIVERSITY 


BY 

EMILY  CROMWELL  SEAMAN,  B.S.,  M.A. 

NEW  YORK  CITY. 


Easton,  Pa.: 

EscHENBACH  Printing  Co. 

1912 


Biochemical  Studies  of  Beryllium 
Sulfate. 


DISSERTATION 


SUBMITTED    IN   PARTIAL   FULFILMENT   OF   THE    REQUIRE- 
MENTS  FOR  THE   DEGREE   OF    DOCTOR   OF   PHIL- 
OSOPHY IN  THE  FACULTY  OF  PURE   SCIENCE 
OF   COLUMBIA   UNIVERSITY 


BY 

EMILY  CROMWELL  SEAMAN,  B.S.,  M.A. 

NEW  YORK  CITY. 


Easton,  Pa.: 

EscHENBACH  Printing  Co. 

1912 


ACKNOWLEDGMENT. 

This  research  has  been  done  at  the  suggestion  and  under 
the  personal  direction  of  Professor  William  J.  Gies.  During 
its  progress  he  has  given  the  investigation  much  personal 
attention.  I  wish  to  express  my  sincere  appreciation  and 
gratitude  for  his  interest  and  assistance  and  especially  I  wish 
to  thank  him  for  the  generosity  which  made  it  possible  for 
me  to  work  in  his  laboratory. 

Emily  C.  Seaman. 

Laboratory  of  Biological  Chemistry  of  Columbia  University, 

College  of  Physicians  and  Surgeons,  New  York  City. 

May  16.  1912. 


CONTENTS. 


Page 

I.  Introduction 7 

II.  General  Influences  on  Nutrition 7 

III.  Pharmacological  Effects 28 

a.  Administration  per  os :^t, 

b.  Subcutaneous  injection  (as  beryllium  sulfate) :^^ 

c.  Subcutaneous  injection  (as  beryllium  lactate) 35 

d.  Intravenous  injection 36 

IV.  Effects  on  Enzyme  Action 41 

a.  Sucrase 41 

b.  Ptyalin 42 

c.  Pepsin 43 

d.  Trypsin 44 

V.  Precipitation  of  Protein 45 

a.  Egg  Albumin 45 

b.  Proteoses 45 

c.  Gelatin 45 

d.  Acid  Meta-protein 45 

VI.  Effect  on  Growth  of  Seedlings 46 

VII.  Effect  on  Bacterial  Growth 49 

VIII.  Summary  of  Conclusions 50 

IX.  Biographical 52 

X.   Bibliography 53 


feces.  The  raw  meat  was  preser\,^ed  in  a  frozen  condition.^ 
The  cracker  meal  was  thoroughly  mixed  and  kept  entirely 
dry  in  large  glass-stoppered  bottles.  The  best  quality  of 
lard  was  purchased  in  small  amounts  and  kept  in  a  re- 
frigerator. Infusorial  earth  Avas  used  instead  of  bone-ash 
in  some  of  the  experiments  in  order  to  prevent  possible  con- 
version of  administered  beryllium  into  the  insoluble  com- 
pounds which  bone-ash  might  induce.  Distilled  water  was 
used.  The  dog  was  given  all  the  daily  portion  of  food  at 
one  time,  the  solid  ingredients  being  thoroughly  mixed  with 
water. 

Periods  and  Weights. — Each  day  of  the  experiment 
ended  at  2.30  p.:m.  The  dogs  were  weighed  just  before 
being  fed.  A  new  period  of  record  was  always  begun  with 
the  day  on  which  the  dog  was  placed  under  new  conditions. 

Collection  of  Excreta. — The  lu-ine  was  collected  each 
day  just  before  feeding,  and  the  daily  volume  recorded. 
The  cage  was  washed  ever}*  second  day,  the  washings  being 
added  to  the  total  urine  for  the  two  days  and  the  volume 
made  up  to  2000  cc.  with  distilled  water.  The  total  nitrogen 
was  determined  from  this.  The  fecal  matter  was  removed 
from  the  cage  as  soon  as  possible  after  its  elimination,  placed 
in  an  evaporating  dish  and  dried  as  speedily  as  possible 
by  exposure  to  the  air.  The  feces  for  each  day  were  kept 
separate  and  weighed  when  thoroughly  dry.  It  was  notice- 
able that,  with  the  exception  of  a  condition  which  will  be  men- 
tioned later  in  the  experiment,  the  appearance  of  the  material 
was  uniformly  dry  and  chalky,  which  agrees  with  the  pre- 
vious experiences  in  the  use  of  infusorialareth  in  nutritional 
experiments  directed  here  by  Prof.  Gies. 

Analysis. — The  total  ingested  nitrogen,  sulfur,  phosphorus 
and  inorganic  matter  were  determined  from  the  food.  The 
nitrogen  of  the  urine  was  determined  on  the  total  volume  for 
every  two  days  and  the  nitrogen  in  the  feces  for  each  period. 
The  ash,  sulfur  and  phosphorus  of  the  urine  and  feces  were  de- 

^  Gies:  American  Journal  of  Physiology,   1901,   v,  p.   235;   Gies  and 
collaborators:  Biochemical  Researches,  1903,  p.  69  (Reprint  No.  i). 


termined  for  the  period.  The  urine  was  preserved  with 
thymol.     The  feces   were  finely  powdered  before     analysis. 

Analytical  Methods. — The  total  nitrogen  was  deter- 
mined by  the  Kjeldahl  process,  oxidation  being  effected 
with  concentrated  sulphuric  acid  and  a  little  cupric  sulfate. 
The  total  sulfur  and  phosphorus  of  the  urine  were  determined 
by  the  caustic  alkali  fusion  methods.  Because  of  the  pres- 
ence of  silicate  in  the  feces  it  was  necessary  to  remove  silica 
after  fusion  before  determining  the  sulfur  and  phosphorus. 
This  was  done  by  dehydration  with  acid  followed  by  desic- 
cation, extraction  and  filtration.  The  total  inorganic  mate- 
rial was  obtained  by  evaporating  the  sample  to  dryness  and 
charring  at  a  low  redness  until  all  the  carbonaceous  material 
had  been  burned  and  only  a  white  residue  remained. 

Beryllium  Sulfate. — The  beryllium  sulfate  used  in  the 
metabolism  experiments  was  a  ptu-e  product  obtained  from 
Dr.  Charles  L.  Parsons  for  this  special  work. 

First  Metabolism  Experiment. — The  dog  selected  for 
this  experiment  was  a  healthy  female  bull  terrier  weighing 
about  twelve  kilos. 

Diet. — The  diet  was  the  same  throughout  the  experiment 
except  that  bone-ash  was  used  during  the  fore  and  first 
dosage  periods.  This  was  then  changed  and  infusorial 
earth  used  in  its  place  for  the  remainder  of  the  metabolism 
work,  for  the  reason  stated  above  (Table  I) . 


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Preparatory  Period. — Bight  days  were  sufficient  for 
the  preparatory  period.  During  this  time  the  animal  became 
accustomed  to  the  food  and  environment.  The  weight 
became  nearly  constant.  The  preparatory  period  ended 
on  Nov.  24,  19 10  at  2.30  o'clock  and  on  that  day  the  experi- 
mental work  was  begun.  It  was  continued  uninterruptedly 
until  2.30  o'clock  on  December  29,  19 10 — (36  days).  The 
experiment  was  divided  into  six  periods  of  different  lengths 
and  conditions  as  follows: 

Fore  Period. — Normal  conditions.  Days,  1-8.  Nov.  24 
to  Dec.  I,  1 9 10.  During  this  period  entirely  normal  condi- 
tions were  maintained. 

First  Dosage  Period. — Influence  of  beryllium  sulfate 
administered  per  os.  Days,  9-16.  Dec.  2-9,  1910.  On  the 
ninth  day  the  first  dose  of  beryllium  sulfate  was  given.  The 
dosage  was  started  with  o.i  gram  and  increased  each 
day  until  on  the  last  day  of  this  period  5.0  grams  had  been 
administered.  The  dose  was  always  given  just  before  feeding 
by  enclosing  the  beryllium  sulfate  in  a  ball  made  of  part  of 
the  weighed  meat.  The  mass  was  swallowed  quickly  without 
mastication  and  the  remainder  of  the  food  given  at  once. 
No  symptoms  were  produced  by  this  amount  of  beryllium 
sulfate  but  it  was  decided  not  to  prolong  this  dosage  period. 
There  was  a  slight  decrease  in  weight. 

Special  Period. — Days,  17-18.  Dec.  lo-ii,  1910.  Be- 
cause of  a  continued  loss  in  weight  and  variations  in 
the  volume  of  urine  it  was  decided  to  consider  the  two  days 
following  the  dosage  period  as  a  special  period. 

Intermediate  Period.  —  Days,  19-24.  Dec.  12-17,  1910. 
During  this  period  there  was  a  tendency  toward  restoration  of 
normal  nutrition.  The  weight  still  decreased  but  toward 
the  end  of  the  period  it  began  to  increase  and  the  volume 
of  urine  struck  a  normal  average. 

Second  Dosage  Period. — Days,  25-28.  Dec.  18-21, 
1 9 10.  After  an  interval  of  eight  days  beryllium  sulfate  was 
again  administered.  As  in  the  first  dosage  period  the  amount 
was  started  with  o.i  gram.     It  was  increased  each  day  until 


by  the  fourth  day,  3.6  grains  of  beryllium  sulfate  had  been 
given.  At  this  point  marked  effects  were  produced.  Previous 
to  the  time  this  amount  was  given  (2  p.m.,  Dec.  21)  there 
were  no  signs  of  sickness.  At  9  a.m.  the  next  morning  (Dec. 
22)  a  large  amount  of  vomit  consisting  of  undigested  meat 
and  mucus  was  found  on  the  pan  and  in  the  urine.  Some  of  the 
material  on  the  pan  was  dry  indicating  that  there  had  been 
an  attack  of  vomiting  sometime  between  midnight  when 
the  dog  was  last  inspected  and  9  a.m.  There  were  no  other 
signs  of  sickness.  The  dog's  nose  was  cold  and  she  seemed 
full  of  life.  The  urine  containing  the  vomit  was  strained 
and  the  solid  particles  from  this  and  from  the  pan  were 
weighed  and  added  to  the  next  day's  food  (Dec.  22).  In 
anticipation  of  vomiting  the  urine  receiver  had  purposely 
been  kept  free  from  thymol.  The  volume  of  urine  was  de- 
cidedly increased,  but  this  was  probably  due  to  the  presence 
of  the  vomit.  Just  before  the  time  for  feeding  on  the  same 
day  the  dog  vomited  twice.  This  vomit  consisted  of  about 
25  cc.  of  colorless  froth  with  a  little  liquid.  It  was  allowed 
to  remain  on  the  pan  to  be  added  to  the  urine.  No  more 
beryllium  sulfate  was  given.  The  maximum  amount  given 
in  this  experiment  was  evidently  a  toxic  dose  for  this  dog 
and  could  not  be  satisfactorily  exceeded  in  such  work.  The 
beryllium  sulfate  had  probably  exerted  a  deleterious  effect 
on  digestion  and  was  possibly  diuretic.  The  feces  for  this 
period  were  black  and  soft  in  marked  contrast  to  those  of  the 
former  periods. 

After  Period. — Days,  29-36.  Dec.  22-29,  19 10.  Although 
no  more  beryllium  sulfate  was  given,  the  dog  showed 
a  tendency  to  vomit  on  the  first  day  of  this  period  (Dec.  23). 
At  II  A.M.  and  again  at  12  m.  she  ejected  small  quantities 
of  thin  liquid.  There  were  no  solid  particles  in  the  vomit 
at  either  time.  The  material  was  added  to  the  day's  urine. 
The  dog  was  very  lively,  the  nose  was  cold  and  she  showed 
no  other  signs  of  sickness.  During  this  after  period  there 
was  a  gradual  but  constant  increase  in  weight. 

Analytical   Results. — The  data  in  the  daily   records  and 


13 

the  summary  of  results  with  the  daily  averages  for  the  first 
metabolism  experiment  are  given  in  Tables  II-IV. 

Discussion  of  Results. — During  the  fore  period  the 
animal  showed  a  fluctuation  in  weight  ending  in  a  slight 
gain.  The  period  was  marked  by  a  decided  plus  balance  of 
the  nitrogen,  ash  and  phosphorus  and  a  small  plus  balance  of 
the  sulfur.  It  was  evident  that  the  diet  was  more  than  suffi- 
cient for  the  dog's  requirements.  The  first  dosage  period 
was  without  marked  effect  although  a  total  of  over  5  grams 
of  beryllium  sulfate  were  given.  The  weight  fluctuated 
slightly  during  the  period  but  there  was  very  little  loss  at 
the  end.  There  was  decrease  of  nitrogen  and  sulfur  elimi- 
nation but  an  increase  in  elimination  of  ash.  The  phos- 
phorus remained  practically  the  same.  There  was  an  in- 
crease in  the  volume  of  urine. 

It  seemed  quite  evident  from  the  data  for  the  two  periods 
following  (and  later  experiments  bear  this  out)  that  the 
beryllium  sulfate  was  cumulative  in  its  influence.  During  this 
time  there  was  a  decided  loss  of  weight  and  further  increase 
in  the  volume  of  urine.  The  first  two  days  were  so  marked 
in  these  respects  that  the  excreta  were  analyzed  for  a  separate 
period.  The  results  showed  a  great  loss  in  nitrogen,  ash, 
sulfur  and  phosphorus  suggesting  that  the  beryllium  sulfate 
had  produced  decided  nutritive  distiu-bance.  During  the 
following  six  days  (the  intermediate  period)  there  was  some 
return  to  normal  conditions  as  was  shown  by  the  decreased 
elimination  of  nitrogen,  sulfur  and  phosphorus. 

In  the  second  dosage  period  nearly  4  grams  of  beryllium 
sulfate  were  given  in  four  days.  It  had  marked  toxic  effect, 
producing  vomiting.  Up  to  this  time  the  feces  had  been 
hard,  chalky,  and  uniform  in  character,  but  on  the  third 
day  of  the  period,  they  began  to  be  dark  and  decidedly  soft. 
This  condition  lasted  until  the  second  day  of  the  after  period. 
There  was  a  marked  loss  of  weight  and  an  increased  elimi- 
nation of  nitrogen  and  sulfur  during  this  second  dosage  period 
but  as  in  the  former  dosage  and  after  periods,  the 
effect  of  the  beryllium  sulfate  extended  beyond  the  time   of 


14 

the  administration.    Here  there  was  a  still  greater  loss  of  body 
nitrogen,  sulfur  and  phosphorus. 

The  result  of  the  first  experiment  showed  a  decided  loss 


Date. 

Nov. 

24 
25 
26 

27 

28 
29 

30 

Dec. 

I 


Body 
weight. 
Kilos. 

11.82 

11.82 

11.82 
11.80 

11.84 
11.84 

11.84 
11.83 


Tabl,e;  II.    Experiment  I. 
Fore   Period. 

Urine. 


BeSO* 
Gram 


Vol. 
cc. 

437 

431 
445 

435 
405 
510 

435 


Reaction, 

Sp.  gr.  litmus. 

1017  acid 

1015  acid 

1016  am.  ac* 
1015  acid 

1015  acid 

1015  acid 

1014  acid 

1015  acid 


Nitrogen. 
Grams. 


12.07 


12.  Il 


11.55 


12.53 


Feces. 
N     Dry 
weight. 
Grams. 

15.4 
31.7 

26.6 
20.2 

14.2 

40. 5 

none 
37.0 


First  Beryllium  Sulfate  Period. 


2 
3 

11.83 
1 1 .  76 

0.1165 
0. 2131 

470 

483 

1015 
1014 

am.  ac. 
am.  ac. 

II 

22 

5-5 
36.0 

4 
5 

11.78 
11.84 

0.2131 
0.2581 

470 
475 

1015 
1015 

acid 
acid 

12 

87 

lost 
17.3 

6 

7 

11.78 
11.80 

05034 
I. 0012 

433 
506 

1014 
1016 

acid 
acid 

10 

03 

20.2 
16.5 

8 
9 

11.79 
11.80 

I . 2473 
I .5002 

483 
418 

1016 
1017 

acid 
acid 

12 

42 

17.0 
350 

10 
II 


11.79 

II  .69 


special  Period. 

480     1017     acid 
550     1015     am.  ac. 


16.16     ^3.5 
14.5 


*  Urines  that  were  amphoteric  but  apparently  more  acid  than  alkalin 
are  referred  to  as  "am.  ac."  in  the  Tables. 


15 
Table  H — (Continued). 

Intermediate  Period. 


Date. 

Nov. 

12 
13 

14 
15 
16 
17 


Body 
weight. 
Kilos. 


Urine. 


71 

68 

69 
69 

73 
70 


BeSOi. 
Grams. 


Vol. 
cc. 

423 
425 

475 
470 
445 


Sp.  gr. 
IOI6 
IOI6 

IOI5 
IOI7 

IOI5 
IOI5 


Reaction, 
litmus. 

acid 
acid 

acid 
am.  ac. 

acid 
acid 


Nitrogen. 
Grams. 


13  59 
13  03 
12.03 


Feces. 

Dry 

wei^t. 

Grams. 

25 -5 
14.0 

20.0 
II  5 

14  5 
17.0 


Second  Beryllium  Sulfate  Period. 


18 

II 

75 

0 

1003 

464 

1015 

am. 

ac. 

19 

II 

74 

0 

5039 

425 

1016 

am. 

ac. 

20 

II 

77 

I 

Olio 

473 

1018 

am. 

ac. 

21 

II 

61 

2 

0100 

595 

1016 

am. 

ac. 

13  32 


14.27 


10.5 

235 

II  5 

18.5 


22 
23 

24 
25 
26 
27 
28 
29 


63 
68 

73 
74 

77 
77 
80 
82 


[fter 

Period. 

415 
446 

1020 
1017 

am. 
am. 

ac. 
ac. 

15 

00 

21  5 
II  5 

433 
490 

1017 
1016 

am. 
am. 

ac. 
ac. 

14 

21 

22.0 
14.0 

455 
455 

1019 
1019 

am. 
am. 

ac. 
ac. 

14 

83 

19  5 

20.0 

415 
468 

1016 
1017 

am. 
am. 

ac. 
ac. 

14 

27 

8.0 
13.0 

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19 


of  weight  accompanied  by  an  increasing  and  prolonged  loss  of 
nitrogen,  inorganic  matter,  sulfur  and  phosphorus  as  the  re- 
sult of  the  disturbance  of  nutrition  which  the  beryllium  sulfate 
induced.    In  this  experiment  the  salt  was  diuretic  in  its  effect. 

Second  Metabolism  Experiment.— The  dog  used  in 
this  experiment  was  the  one  employed  in  the  first.  Although 
she  seemed  in  normal  condition  at  the  end  of  the  last  ex- 
periment a  month  elapsed  before  the  second  one  was  begun. 
During  this  time  the  dog  appeared  to  be  perfectly  healthy 
and  was  very  lively. 

Diet. — The  character  of  the  diet  was  the  same  as  that  in 
the  first  experiment. 

Table  V. 


Composition  of  Daily  Diet 

Lean 
Ingredients.  meat. 

Days  1-24 

Grams 


Cracker 
meal. 


Daily  amount    178 
Nitrogen  6 .  69 

Sulfur 
Phosphorus 
Inorganic  mat- 
ter I 


1-24 

Grams. 

48 
I  .06 


Lard. 
1-24 

Grams 

35 
0.0002  0.00012 


Experiment  11. 

Infusorial 
earth. 

1-24 

Grams. 
10 


Water. 
1-24 


415 


0.338   0.078 
0.16      0.06 

96     O . 62 


9  7 


Total. 
Grains. 

686 

7-75 
0.42 
0.22 

3-45 


Fore  Period. — Normal  conditions.  Days,  1-8.  Jan.  27 
to  Feb.  3,  1911.  During  this  period  there  was  a  slight  varia- 
tion in  the  amount  of  daily  urine  and  a  small  gain  in  weight. 

Dosage  Period. — Days,  9-16.  Feb.  4-11,  1911.  The 
dosage  period  began  at  2.30  o'clock  on  Feb.  4.  The  amount 
of  beryllium  sulfate  given  was  0.5  gram.  It  was  increased 
half  a  gram  each  day  until  on  Feb.  6,1.5  gram  were  given.  On 
the  following  morning  at  9  o'clock  vomit  was  found  in  the  cage. 
It  consisted  of  a  few  particles  of  undigested  meat  in  a  mass 
of  brownish  mucus.  The  vomit  amounted  to  8  grams. 
This  was  added  to  the  food  for  Feb.  7.  No  beryllium  sulfate 
was  given  that  day.  The  following  day  (Feb.  8)  the  dog 
appeared  to  be  normal.  The  dose  was  again  omitted  but 
on  Feb.  9  the  dog  was  given  i  gram  of  beryllium  sulfate. 
On  the  following  morning  the  dog  vomited.  The  vomit 
consisted  of  mucus  which  was  added  to  the  food.     On  Feb. 


20 


lo  the  dose  was  omitted;  the  dog  was  not  sick.  On  Feb. 
II,  I  gram  of  beryllium  sulfate  was  given.  The  dog  was 
sick  the  following  morning  but  only  to  a  slight  degree — less 
so  than  at  any  time  previous.  It  was  decided  to  close  the 
dosage  period  at  this  point  as  it  was  evident  that  the  per- 
missible maximum  dose  had  been  given. 

After  Period. — Days,  17-24.  Feb.  12-19,  1911.  The 
period  consisted  of  eight  days  during  which  time  no  beryllium 
sulfate  was  given.  The  data  for  the  experiment  are  given 
in  Tables  VI-VIII. 

TabIvE  VI.    Experiment  II. 
Fore  Period. 


Body 
weight. 
Kilos 

BeSO*. 
Gram. 

Urine. 

Feces. 

Dry 

weight. 

Grams. 

Date. 
Jan. 

Vol. 
cc. 

Sp.  gr. 

Reactions, 
litm^us. 

Nitrogen. 
Grams. 

27 

[2.23 

470 

IO16 

acid 

25.0 

28 

[2.30 

410 

IO16 

acid 

13 -43 

12.0 

29 
30       ] 

[2.36 
[2.36 

450 
490 

IO16 
IO16 

acid 
acid 

14.38 

9.0 
23.0 

31 
Feb. 

I        ] 

[2.37 
[2.32 

540 
490 

IOI5 
IOI7 

acid 
acid 

15.22 

16.0 
13.0 

2        ] 

t2.34 

510 

IOI5 

acid 

8.0 

3      J 

[2.34 

485 

IOI6 

acid 

1 1  .  71 

24.0 

Beryllium 

Sulfate  Period. 

4      ] 

[2.33 

0.5108 

500 

1016 

acid 

14.56 

16.5 

5      1 

[2.31 

I . 0078 

510 

1016 

acid 

II-5 

6     ] 

12.30 

1-5117 

450 

1020 

am.  ac. 

14.48 

25-3 

7     ] 

[2.28 

485 

1018 

acid 

10.5 

8      ] 

[2  .  29 

430 

1020 

acid 

19-5 

9      ] 

12.33 

I . 0200 

495 

1017 

acid 

1557 

none 

10     ] 

[2.36 

390 

1018 

acid 

20.0 

II      ] 

[2.32 

I .0114 

555 

1017 

acid 

15-51 

27-3 

After  Period. 

12  ] 

13  1 

[2.27 
2.32 

540 
460 

1017 
1018 

acid 
acid 

15.00 

8.4 
9-5 

14     ] 

2.32 

445 

1019 

acid 

19. 1 

15      1 

2.31 

510 

1017 

acid 

15-57 

19.0 

16  ] 

17  ] 

[2.22 
12.29 

550 
350 

1017 
102 1 

am.  ac. 
am.  ac. 

15.68 

lO.O 

9.0 

18      ] 

[2.38 

415 

1017 

acid 

27.2 

19     ] 

t2.34 

535 

1017 

acid 

15.40 

5-2 

21 


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23 

Discussion  of  Results. — The  fore  period  showed  that 
the  animal  continued  to  gain  weight  on  the  prescribed  diet 
as  in  the  former  experiment.  During  the  period  in  which 
5  grams  of  beryllium  sulfate  were  administered  there  was  a 
fluctuation  in  weight  which  continued  into  the  after  period 
and  a  slight  decrease  in  the  total  volume  of  urine.  The  daily 
volume  varied  considerably  but  these  variations  usually  at- 
tended pronounced  sickness  as  the  result  of  the  dosage. 
The  dosage  period  showed  a  marked  increase  in  the  elimina- 
tion of  nitrogen  and  phosphorus,  but  the  sulfur  and  total  in- 
organic matter  were  not  much  affected. 

The  after  period  showed  a  continued  and  increased  elimi- 
nation of  body  nitrogen.  The  weight  of  the  animal  fluctuated 
during  this  time  but  by  the  end  of  the  period  there  was  a  re- 
turn to  normal.  There  was  an  increased  elimination  of  sulfur 
but  less  phosphorus  than  in  the  dosage  period.  The  data  for 
the  analysis  showed  the  same  nutritive  disturbances  as  in 
the  first  experiment  although  not  to  so  marked  a  degree. 

Third  Metabolism  Experiment. — The  animal  used  in 
this  experiment  was  a  medium  sized  short  haired  dog  weighing 
nearly  lo  kilos. 

Diet. — The  diet  was  of  the  same  character  as  in  the  two 
former  experiments. 


Table  IX. 
Composition  of  Daily  Diet.   Experiment  III. 


Lean 

Cracker 

Infusorial 

Ingredients. 

meat. 

meal.          Lard. 

earth. 

Water.     Total. 

Days 

I-18 

I-18          I-18 

I-18 

I-18 

Grams. 

Gram.        Grams. 

Grams. 

cc.         Grams. 

Daily  Amount 

160.0 

0.40   30.0 

7.0 

350      587    0 

Nitrogen 

6.4 

0.88      0.0018 

7-3 

Sulfur 

0.30 

0.06 

0.36 

Phosphorus 

0.  16 

0.05 

0.21 

Inorganic    mat- 

ter 

1.6 

0.32 

6.79 

8.71 

24 

Preparatory  Period. — This  period  consisted  of  15  days. 
It  was  two  or  three  days  before  the  dog  became  accustomed  to 
its  new  environment  and  ate  all  the  daily  amount  of  food. 
During  this  time  there  was  great  fluctuation  in  the  amount 
of  daily  urine.  On  the  last  day  of  the  period  there  was  com- 
plete retention  of  the  urine  and  the  large  volume  on  the  first 
day  of  the  fore  period  was  the  extra  elimination. 

Fore  Period. — Normal  condition.  Days,  1-6.  May  16- 
21,  191 1.  With  the  exception  of  the  fluctuation  in  the  daily 
volume  of  urine  normal  conditions  were  maintained  through- 
out the  period. 

Dosage  Period.  —  Days,  7-12.  May  22-27,  1911.  On 
the  7th  day  beryllium  sulfate  was  given  per  os  just  before 
feeding  as  in  the  previous  experiments.  The  dosage  was 
started  with  0.5  gram  and  gradually  increased  until  on  the 
third  day  of  the  period  the  dog  received  i  gram.  This  was 
followed  by  very  slight  vomiting.  The  same  dose  was  re- 
peated on  the  following  day.  Slight  vomiting  resulted  the 
next  day  at  noon.  This  was  frothy  and  unformed.  On 
each  of  the  two  days  following  the  dog  received  i  gram  of 
the  sulfate  with  the  food.  The  vomit  as  a  result  each  time 
was  stringy  and  mucous-like.  The  decided  increase  in 
amount  showed  that  the  dog  was  at  the  point  of  very  marked 
effect  from  the  beryllium  sulfate  and  the  dosage  was  stopped. 

After  Period. — Days,  12-18.  May  28  to  June  2,  191 1. 
During  this  period  normal  conditions  were  maintained. 

Analytical  Results. — The  analytic  data  pertaining  to  this 
experiment  are  recorded  in  Tables  X-XII. 


25 


Date. 
May. 

i6 

17 
18 

19 
20 
21 


Body 
weight. 
KUos. 

9.60 

9.68 

9.70 

9-73 
9-75 
9- 63 


BeSOi. 
Gram. 


Table  X. 

Experiment  III. 

Fore  Period. 

Urine. 


Vol. 
cc. 

475 
250 
320 
355 
355 
455 


Reaction, 
Sp.  gr.    litmus. 


IOI9 
IOI5 
1020 
IOI7 
IO17 
1020 


acid 
acid 
acid 
acid 
acid 
acid 


Nitrogen. 
Grams 


13    05 

12.86 
14.2 


Feces. 

Dry 
weight. 
Grams. 

7.0 

none 
12.2 
8.4 
10.6 
19.2 


Beryllium  Sulfate  Period. 


22 

9 

23 

9- 

24 

9 

25 

9- 

26 

9- 

27 

9 

.65  0.5049  310  1020  acid 

.66  0.7543  300  1 02 1  acid 

.72  1.0055  340  1017  acid 

.65  1.0293  440  1019  acid 

.65  0.9890  400  1020  acid 

.75  1. 0000  275  loio  acid 


12.6 
143 
154 


9 
II 

9 
II 

8 
13 


After  Period. 


28  9.65 

29  9.68 

30  9.64 

31  9.69 

June 

1  9.86 

2  9.68 


525  102 1  acid 

325  1015  acid 

390  1020  acid 

325  1020  acid 

220  1 05 1  acid 

490  1020  acid 


15.0 


14  3 


143 


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28 

Discussion  of  Results. — With  this  animal  the  4  grams 
of  beryllium  sulfate  produced  little  variation  in  weight  and 
total  volume  of  urine  for  the  period.  The  results  of  the 
analysis,  however,  show  the  same  effects  on  nutrition  as  were 
produced  in  the  two  former  experiments.  The  eliminations 
of  body  nitrogen,  total  inorganic  matter,  sulfur  and  phos- 
phorus were  affected,  extending  beyond  the  dosage  period. 
The  increased  loss  of  nitrogen  manifested  itself  more  in  the 
after  period. 

The  results  obtained  in  the  third  experiment  corroborated 
those  of  the  first  and  second. 

General  Remarks. — From  the  three  experiments  performed 
to  ascertain  the  effect  of  beryllium  sulfate  on  metabolism,  it 
was  clearly  shown  that  the  salt  produced  pronounced  nutritive 
disturbances.  It  was  demonstrated  in  the  fore  period  of  each 
experiment  that  the  daily  diet  was  more  than  sufficient  for 
the  animal  and  the  disturbed  metabolism  which  developed 
during  the  dosage  and  after  periods  must  have  been  due  to  the 
direct  influence  of  the  beryllium  sulfate. 

The  metabolic  disturbance  was  shown  before  sufficient 
amounts  of  the  salt  were  given  to  produce  vomiting  as  was 
illustrated  in  the  first  dosage  period  of  the  first  experiment. 

III.     PharmacoivOgical  Effects.^ 

Some  earlier  workers  in  endeavoring  to  establish  the  atomic 
weight  of  beryllium  and  its  place  in  the  periodic  system 
made  a  few  comparative  studies  of  the  pharmacological 
effect  of  salts  of  beryllium,  aluminium,  barium,  strontium  and 
calcium.  As  early  as  1882,  Blake  asserted  that  the  general 
physiological  effects  of  beryllium  were  similar  to  those  of 
aluminium.^ 

*  The  pharmacological  experiments  are  in  progress  at  this  writing 
(May  14,  1912). 

^  Blake:  "Atomic  Weights  of  Beryllium  as  Determined  by  its  Physio- 
logical Actions."  Chemical  News,  \xv,  p.  III.  Ueber  den  Zusammenhang 
der  molekularen  Eigenschaften  anorganischen  Verbindungen  und  ihre  Wirk- 
ung  auf  den  lebenden  thierischen  Organismus.  Berichte  der  deut.  chem. 
Gesellschaft,  xiv,  p.  394. 


29 

He  used  the  sulfate  salt  and  found  that  0.023  gram  per 
kilo  of  body  weight  was  a  lethal  dose.  His  experiments 
were  performed  on  dogs  but  he  does  not  describe  the  symp- 
toms produced. 

In  1884,  Brunton  and  Cash^  in  studying  the  comparative 
toxicity  of  the  alkali  earths  found  that  irritation  of  the 
spinal  cord  was  marked  in  cases  of  poisoning  by  beryllium. 
Beryllium,  calcium,  strontium  and  barium  all  paralyzed  the 
motor  nerves  to  some  extent.  These  workers  used  beryllium 
chloride  in  their  experiments  and  performed  them  on  frogs. 
They  found  that  the  substance  acts  as  a  poison  with  0.0013 
gram  of  BeClj  to  i  gram  of  body  weight  of  frog.  The  table 
on  the  following  page  is  a  summary  of  their  results. 

In  a  study  of  the  comparative  toxicity  of  aluminium  and 
beryllium,  Siem-  found  that  subcutaneous  and  intravenous 
injections  of  beryllium  were  more  toxic  than  those  of  alumin- 
ium. He  used  solutions  of  the  lactate  and  tartrate  salts_ 
of  beryllium  and  performed  his  experiments  on  frogs,  cats, 
dogs  and  rabbits.  He  calculated  his  doses  in  terms  of  BeO 
and  states  that  from  0.02-0.028  gram  BeO  was  a  lethal  dose 
for  a  medium  sized  frog;  from  0.004-0.005  gram  BeO  per 
kilo  of  body  weight  for  dogs  and  cats  and  from  0.008-0.01 
gram  per  kilo  of  body  weight  for  rabbits.  Only  in  one  ex- 
periment with  a  frog  in  which  the  injection  was  made  in  the 
dorsal  lymph-sac  was  the  dose  without  effect.  In  all  other 
experiments  the  animal  died  in  a  few  days. 

In  the  frogs  he  found  the  effect  of  beryllium  to  be  as  follows : 
A  central  paralysis  due  to  the  direct  effect  on  the  ganglia 
of  the  brain  and  spinal  cord.  The  reflexes  were  impaired 
and  the  breathing  became  irregular.  The  action  of  the  heart 
was  rhythmical  but  grew  gradually  slower.  The  frogs  died 
on  the  day  following  the  injection. 

In  other  animals  the  toxicity  of  the  beryllium  was  ac- 
companied   by    loss    of    appetite    and    impaired    activity  of 

'  Brunton  and  Cash:  Royal  Society,  Philosophical  Transactions, 
1884,  p.  197. 

^vSicm:  "Ueber  die  Wirkung  des  Aluminiums  und  Berylliums  auf 
den  thierischen  Organismus."     Inaug.  Dissert.,  Dorpat,  1886. 


30 


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32 

the  intestines  usually  after  a  single  diarrheal  bloody  dis- 
charge. There  was  vomiting,  marked  physical  depression 
and  rapid  loss  of  body  weight  and  temperature.  At  first 
the  animal  became  lethargic,  lying  on  its  side  and  showing 
great  reluctance  to  move.  When  this  was  done  the  animal 
showed  ataxic  movements.  This  condition  was  followed  by 
general  muscular  tremor  of  short  or  long  duration  accompany- 
ing a  complete  loss  of  sensation.  The  animal  usually  died 
in  a  convulsion.  The  heart  beat  became  weak  but  remained 
rhythmical  and  its  action  outlasted  the  respiration  always 
by  a  minute.  The  motor  nerves  and  muscles  remained  ir- 
ritable up  to  the  time  of  death.  The  amount  of  urine  ex- 
creted was  slight  and  usually  contained  albumin. 

Siem  administered  the  beryllium  either  subcutajieously 
or  intravenously.  Beryllium  tartrate  was  used  for  the 
subcutaneous  injection  and  the  double  salt  of  beryllium  lac- 
tate for  intravenous  injections.  The  animals  usually  died 
on  the  third  day. 

Pathological  examination  showed  the  liver  and  kidneys 
to  be  much  affected.  The  former  was  a  deep  yellow  color, 
the  parenchyma  soft  and  fragile,  without  granular  structure, 
and  bloodless.  A  microscopic  examination  showed  the  cells 
of  the  liver  to  have  been  destroyed  giving  place  to  a 
yellowish  red,  finely  granular  mass.  There  was  also  a  de- 
cided congestion  of  the  alimentary  canal,  especially  in  the 
case  of  the  dogs.  The  entire  mucous  membrane  of  the 
stomach  and  particularly  of  the  small  intestine  was  deep 
red  and  swollen.  The  intestine  showed  marked  ulceration. 
Siem  found  that  the  general  effects  of  beryllium  were  the  same 
as  that  of  aluminium  except  that  the  toxic  effects  of  beryl- 
lium were  more  pronounced. 

In  the  investigation  carried  on  in  this  laboratory  it  was 
decided  to  first  ascertain  the  effects  of  the  beryllium  sulfate 
if  a  calculated  toxic  dose  was  given  per  os.  The  material 
used  in  all  these  experiments  was  a  Kahlbaum  preparation. 

In  the  study  of  its  effect  on  nutrition,  it  was  found  that 
O.I  gram  of  beryllium  sulfate  per  kilo  of  body  weight,  given 


33 

per  OS  produced  sickness  in  a  dog.  This  was  equivalent  to 
0.014  gram  of  BeO. 

First  Pharmacological  Experiment. — Dose  given  per 
OS.  A  short  haired  dog  weighing  6.08  kilos  was  chosen 
for  this  experiment.  The  animal  was  lively  and  in  good 
condition.  Four  times  the  amount  of  the  toxic  dose  was 
administered.  As  this  was  of  considerable  bulk,  it  was 
divided  and  given  in  three  balls  of  meat,  one  immediately 
after  the  other.     Each  one  was  swallowed  without  mastication. 

July  22,  191 1.  11.30  A.M.  2.38  grams  of  BeSO^  given. 
12.30  P.M.  Dog  lying  quietly  in  cage  but  apparently  well. 
Stood  up  without  difficulty. 

1. 00  P.M.     Vomited  most  of  the  meat  in  solid  masses. 

2.30  P.M.     Dog  apparently  well. 

5.30  P.M.     Ate  a  little  food  and  drank  water. 

6.00  P.M.     Dog  seemingly  normal  again. 

Second  Pharmacological  Experiment. — Dose  given  per 
OS.  The  same  animal  was  used  in  this  experiment  as  in  the 
former  one.  Three  days  were  allowed  to  elapse  before  re- 
newing the  dosage.  During  that  time  the  dog  appeared  to 
be  perfectly  normal — eating  and  retaining  all  the  food  given. 
In  this  second  experiment  the  amount  of  beryllium  sulfate 
was  four  times  that  given  in  the  former  one.  The  dose 
was  administered  in  five  balls  of  meat. 

July  25,  191 1.     2.30  p.m.     8.95  grams  of  BeSO^  given. 

3.00  p.m.  Vomited,  apparently  all  of  the  meat.  Frothy 
liquid  in  the  bottom  of  the  cage. 

3.05  P.M.     Vomit,  white  frothy  liquid. 

3.15  P.M.     Dog  quiet.     Stood  in  cage  without  difficulty. 

3.30  P.M.     No  symptoms. 

4.00  P.M.     No  symptoms. 

Conclusion.  From  the  above  results  it  was  apparent  that 
the  beryllium  sulfate  caused  vomiting  before  a  sufficient 
amount  of  the  material  was  absorbed  to  produce  a  fatal  re- 
sult or  even  strong  toxic  symptoms 

Third  Pharmacological  Experiment.  Subcutaneous 
Injection. — Siem  states  that  from  0.004  to  0.005  gram  of  BeO 
per  kilo  of  body  weight  is  a  lethal  dose  for  a  dog.     He  made 


34 

daily  injections  of  small  amounts  until  the  lethal  dose  was 
administered.  In  the  following  experiments  the  whole  dose 
was  given  in  single  injections.  In  the  first  experiment  the 
amount  administered  was  equivalent  to  0.003  gram  of  BeO 
per  kilo — slightly  below  the  lethal  dose  as  determined  by 
Siem.  This  was  done  to  ascertain  what  symptoms  would 
be  produced. 

The  animal  used  in  the  previous  pharmacological  work 
was  used  in  this  experiment.  The  dog  was  in  perfectly  normal 
condition  and  lively.  The  BeS04  was  dissolved  in  the  smallest 
possible  amount  of  physiological  salt  solution  and  the  in- 
jection made  over  the  right  kidney,  the  area  being  previously 
shaved  and  thoroughly  cleansed. 

July  29,  1911.  Dog  weighing  6.05  kilos.  3.45  p.m.  0.14 
gram  of  BeSO^  injected  subcutaneously. 

4.30  P.M.  Dog  lively.  Fed  at  this  time.  All  food  taken 
and  retained.     Feces  hard  and  dry. 

Midnight.  Dog  gradually  becoming  more  reluctant  to 
rise  and  evidently  getting  sick. 

July  30.  10.00  A.M.  Dog  lying  quietly  in  cage,  no  at- 
tempt to  move.  When  lifted  to  her  feet  the  animal  showed 
signs  of  pain.  Breathing  deeper  and  slower.  Shivered 
occasionally.     Nose  warm. 

These  conditions  continued  through  the  morning  but 
during  the  afternoon  they  began  to  disappear.  She  moved 
with  difficulty  but  otherwise  appeared  normal.  The  nose 
was  warm  and  albumin  was  found  in  the  urine.  All  the  food 
was  taken  and  retained.  During  the  following  day  the 
animal  became  more  lively. 

On  the  third  day  following  the  injection  an  edematous 
swelling  developed  under  the  groin  on  the  side  in  which  the 
injection  was  made,  the  fluid  extending  under  the  abdomen. 

August  2.  3.30  P.M.  0.28  gram  of  BeS04  was  given  by 
subcutaneous  injection  over  left  kidney.  No  symptoms 
were  produced  on  the  day  the  injection  was  given.  On  the 
following  morning,  the  dog  was  lying  quietly  in  the  cage  re- 
luctant to  move,  but  otherwise  in  the  same  condition  as  before 
the  administration  of  the  beryllium  sulfate.     The  breathing 


35 

was  natural;  the  edema  produced  by  the  first  injection  had 
become   slightly  less.     On   the   third   day  following   the   in- 
jection oa  the  left  side  an  abscess  developed  at  the  point  of        S\ 
the  second  injection. 

Fourth  Pharmacological  Experiment. — A  second  ex- 
periment with  subcutaneous  injection  was  made  repeating 
the  conditions  of  the  former.     The  dog  weighed  6.8  kilos. 

Aug.  5,  19 II.  11.00  A.M.  0.667  gram  of  BeSO^  was  given 
subcutaneously  over  the  right  kidney.  By  the  afternoon  the 
animal  seemed  reluctant  to  stand  and  had  difficulty  in  moving 
the  right  hind  leg.  There  was  slight  shivering,  deep  breath- 
ing and  considerable  saliva.  Food  was  refused.  Albumin 
appeared  in  the  urine  voided  late  in  the  afternoon. 

Three  days  after  the  administration  of  the  beryllium  sulfate 
decided  gangrene  developed  on  the  side  of  the  injection:  the 
gland  under  the  right  groin  became  swollen.  No  other 
symptoms  developed. 

Aug.  II,  1911.  3  P.M.  1.285  grams  BeSO^  were  injected 
over  left  kidney.  The  same  results  were  obtained.  Necrosis 
extending  over  the  thigh  developed  on  the  side  of  injection  two 
days  after  the  administration  of  the  dose. 

Fifth  Pharmacological  Experiment.  Subcutaneous 
Injection  of  Beryllium  Lactate.  —  Because  of  the  marked 
contrast  in  results  obtained  in  the  above  experiments  and 
those  reported  by  Siem  it  was  decided  at  this  point  to  change 
the  method  employed  to  conditions  more  nearly  those  used 
by  him. 

The  lactate  salt  was  prepared.  This  was  done  by  dissolv- 
ing the  beryllium  sulfate  in  the  smallest  possible  volume  of 
water  making  the  solution  alkaline  with  i  per  cent,  sodium 
hydroxide  and  then  faintly  acid  with  lactic  acid.  This  solu- 
tion was  used  for  the  subcutaneous  injection. 

Aug.  12,1911.  Dog  weighing  5.8  kilos.  12. com.  i. 17  grams 
BeSO^  (as  lactate)  were  injected  subcutaneously  in  right  side. 
No  symptoms  were  produced  until  the  following  day  when  a 
large  swelling  appeared  under  the  right  groin.  The  place 
of  injection  had  become  large  and  soft.  No  other  symptoms 
developed.     The  animal    appeared  otherwise  normal.     There 


36 

was  no  sign  of  pain  in  the  affected  area.  The  dog's  nose 
was  cold. 

Sixth  Pharmacological  ExPiBRiMENT.  Intravenous  In- 
jection.— According  to  Siem^  0.004-0.005  gram  of  BeO  per 
kilo  of  body  weight  is  a  lethal  dose  for  a  dog  or  cat.  In  his 
first  experiment  with  an  intravenous  injection  he  administered 
0.04  gram  BeO  to  a  cat  weighing  2.520  kilos.  The  injection 
was  made  at  ten  o'clock  in  the  morning.  By  noon  of  the  same 
day  the  animal  had  become  apathetic  and  reluctant  to  move. 
It  drank  some  milk  but  this  was  soon  vomited.  On  the 
following  morning  a  general  weakness  had  developed.  The 
cat  was  staggering  and  the  senses  much  affected.  During 
the  night  there  had  been  more  vomiting.  This  consisted  of 
a  white,  frothy  mass.  By  noon  of  this  day  the  senses  were 
practically  deadened.  At  his  time  respiration  was  48,  the 
pulse  220-230  and  very  weak,  the  temperature  34.7°  (98.4 
F.).     On  the  third  day  the  animal  died. 

In  the  second  experiment  Siem  injected  o.i  gram  of  BeO 
into  a  dog  weighing  16.650  kilos  at  three  o'clock  in  the  after- 
noon. During  the  night  there  was  much  vomiting  and  diar- 
rheal feces.  On  the  second  day  the  dog  weighed  15.630 
kilos.  It  staggered  about  and  dragged  the  hind  part  of  the 
body.  When  the  animal  lay  still  on  its  side,  it  showed  ner- 
vous twitching  of  the  muscles.  The  senses  were  decidely 
dulled.  The  pulse  was  100  and  rather  strong;  the  tempera- 
ture 35.8°  (96.4  F.).  On  the  third  day  the  weight  had  de- 
creased to  14.340  kilos;  the  pulse  was  160;  the  dog  lay  per- 
fectly motionless,  the  senses  entirely  deadened.  At  2  o'clock 
the  respiration  was  very  superficial  and  the  animal  died  in 
a  sudden  convulsion. 

In  the  first  experiment  of  this  research  made  with  an  in- 
travenous injection,  the  amount  given  was  a  little  more 
than  the  lethal  dose  prescribed  by  Seim.  30  cc.  of  i  per  cent, 
aqueous  solution  of  beryllium  sulfate  was  administered. 
The  dog  weighed  6.2  kilos,  and  the  amount  injected  equalled 
0.0067   gram    BeO   per   kilos.      The    Kahlbaum   preparation 

^  Loc.  cit. 


37 


of  the  salt  was  used  and  the  solution  made  up  with  distilled 
water.     The  conditions  of  the  experiment  were  as  follows: 
April  4th,  191 2.     Dog  weighing  6.2  kilos. 
3.15  P.M.    Preparation  begun. 

Respiration,  36. 

Pulse,  132. 

Injection  started. 

Injected  i  cc. 

Injected  2  cc;  respiration,  32;  pulse,  132. 

Injected  3.5  cc. 

Respiration,  32. 

Pulse,  140;  injected  5  cc. 

Injected  5.6  cc. ;  injection  stopped. 

Pulse,  144;  irregular. 

Heart  more  regular;  respiration,  28;  slowing. 

Injection  started  again. 

Injected  7  cc. 

Pulse,  142. 

Very  rapid  heart  action.     Injected  8  cc. ;  long 


3-45 
346 
3-47 

3-47-3/4 
350 

3-53-1/2 
3-54 
3-55 
3-56 

3-57-1/2 
3-58 

3-58-1/2 
4.00 
4.1 

4.2-1/2 
inhalation. 

4-3 
4-3-1/2 

4-5 
4.6 

4.8 
given) . 

4. 1 1 

4-12 
increased. 

4.14 


Breathing  very  shallow. 

Respiration,  28 ;  injected  8.2  cc. ;  injection  stopped. 

Heart  action  rapid  and  weak. 

Heart  very  irregular. 

Pulse,    140;    respiration,    32    (o.ooi    gram   BeO 

Injection  started  again. 

Shivering  tendency  at  the  beginning  has  steadily 

Respiration,   26;  irregular — shivering  increasing 


now  affecting  fore  legs. 

4.16  Respiration,  32. 

4.16-1/2  Injected  14  cc;  abdomen  getting  pink;  salivation 
increasing. 

4.21  Respiration,  28;  injected  18  cc. 

4.22  Temperature,  102.6;  injected  i8.6  cc. 

4.34  Tremor  increasing;  extending  over  entire  body. 

4.35  Dog  becoming  uneasy. 


38 

4.38  Respiration,    52;    injected    25    cc. ;    wagged   tail 

when  called;  made  effort  to  get  off  board. 

4.42  Temperature,    102.6;   injected   30   cc.     Injection 

stopped;  respiration,  48;  heart  weak  and  irregular;  pulse,  144. 
Abdomen  continues  pink. 

4.50  Put  in  cage,  wagged  tail,  nose  cold,  apparently 

normal.     Heart  beat  normal,  strong;  respiration,  32. 

5.10  Temperature,  103.4. 

'5.12  Fed.     All  food  taken. 

5.34  Temperature,  102.6. 

7.30  Temperature,  105.2. 

When  the  dog  was  seen  late  in  the  evening  it  had  vomited 
twice.  The  material  was  soft  but  not  liquid.  Since  being 
returned  to  the  cage  275  cc.  of  dark  urine  had  been  passed 
which  contained  a  trace  of  albumin.  The  feces  were  soft. 
The  animal  lay  in  the  corner  of  the  cage  refusing  to  stand. 
The  temperature  was  105°,  the  respiration  28  and  the  pulse 
fast.  At  9  A.M.  on  the  following  day,  the  feces  contained 
bloody  mucous;  the  urine  (175  cc.)  was  light  in  color  and  re- 
sponded very  strongly  to  the  albumin  test.  The  temperature 
was  still  105°;  the  weight  5.68  kilos.  The  dog  stood  when 
lifted  but  walked  about  the  cage  with  difhculty.  During 
the  afternoon  the  dog  became  decidedly  depressed,  refused 
food  but  drank  a  little  water.  At  9  o'clock  in  the  evening 
the  temperature  was  104;  urine  50  cc. 

April  6th,  10  A.M.  Temperature,  100.2.  Vomit  frothy  and 
mucous.  Weight,  5.51 ;  urine  160  cc.  Dog  stood  and  walked 
about  the  cage  more  easily  than  on  previous  day.  Refused 
food  and  water.  Nose  cold.  At  noon  the  dog  tried  to  vomit, 
there  were  repeated  attacks  of  retching  and  much  froth 
about  the  mouth.  The  animal  walked  easily  about  the  cage. 
During  the  day  there  was  increased  depression. 

6.30  P.M.     Pulse,  100;  respiration,  16;  cold. 

8.15  P.M.     Dog  died  in  asphyxia. 

An  autopsy  the  following  day  showed  the  heart  in  diastole ; 
blood  very  "black";  large  intestine  greatly  congested  and 
•small  intestine  practically  empty  and  flabby;  stomach  con- 
tained bloody,  thin  fluid.     The  bile  was  thick  and  black;  the 


39 

lungs,  liver,  kidneys,  pancreas,  and  spleen  normal.  The 
bladder  was  empty. 

Before  an  experiment  was  tried  on  another  dog,  the  new 
animal  was  kept  under  observation  for  a  week  and  the  daily 
volume  of  urine,  weight  of  the  dog  and  character  of  the  feces 
recorded.  The  second  experiment  was  performed  under 
the  same  conditions  but  half  the  amount  reported  by  Siem 
as  the  lethal  dose  was  injected.  16.4  cc.  of  a  i  per  cent, 
solution  of  beryllium  sulfate  was  injected  into  a  dog  weighing 
8.89  kilos  which  equaled  0.0025  gram  BeO  per  kilo. 

The  conditions  of  the  experiment  were  as  follows: 

April  18,  1912.     3.15  P.M.     Operation  begun. 

3.36  Pulse,  138;  respiration,  28. 

3.38  Pulse,  134;  respiration,  36. 

3.40  Pulse,  128. 

3.42-1/2    Respiration,  32;  pulse,  128. 

3.43  Injection  begun. 

3.45  Pulse,  120;  somewhat  irregular. 

3.46  Injected  3  cc. 

3.47  Pulse,  108;  irregular. 

3.48  Respiration,  28;  injected  6  cc;  dog  trembling. 

4.51  Injected  11  cc.  Muscular  trembling  increased; 
heart  action  weaker. 

4.53  Respiration,    24.     Tremor    in    muscles    prevents 

getting  pulse;  abdominal  surface  warm  and  pink;  nose  cold. 
4.55  Injected  14  cc. 

4.58  Injected  17  cc.  Difficulty  in  breathing;  heart 
beat  cannot  be  felt  on  chest. 

4.59  Injected  18  cc.  (by  correction  for  amount  left 
in  tubing  16.4  cc). 

5.08  Heart  very  weak. 

5.10  Experiment  finished. 

5.1 1  Weight  9.26  kilos;  dog  in  cage. 

5.52  Respiration,  32;  pulse,  116;  temperature,  102. i. 
6.10  Diarrheal    feces.      100   cc.     urine,     no    albumin. 
April  19.  8.45  A.M.    Temperature,  105.7;  pulse,  144;  urine, 

330  cc.  dark,  no  albumin,  feces  soft  but  normal.  Food 
taken  and  retained. 


40 

12  M.  Temperature,  105;  pulse,  136. 

I  P.M.  Temperature,  105.4;  pulse,  140;  urine,  50  cc. 
albumin. 

3.15  P.M.   Nose  warm  and  dry.     Dog  markedly  less  lively. 

5.30  P.M.  Temperature,    105.7;  pulse,    124;  irregular. 

7.00  P.M.    Urine,  50  cc,  light,  no  albumin,  feces  hard. 

April  20,  8  A.M.  Urine,  80  cc,  slight  albumin,  reduction; 
pulse,  148;  regular;  temperature  104.7,  the  animal  showed 
occasional  muscular  trembling.  All  food  and  water  refused. 
These  same  conditions  continued  through  the  day  except 
that  during  the  afternoon  the  temperature  dropped  to  103.6. 

April  21,  10  A.M.  There  had  been  bleeding  from  the  dog's 
wound  during  the  night.  The  animal  was  decidedly  sick 
and  refused  all  food.  There  was  considerable  frothy  vomit 
on  the  pan  and  bloody,  diarrheal  feces. 

4.00  P.M.    Very  little  sign  of  life.     Animal  perfectly  quiet. 

4.10  P.M.  Animal  dead. 

The  autopsy  the  following  day  showed  the  heart  in  diastole 
and  full  of  blood;  lungs  congested;  blood  "black";  spleen 
large  and  black  along  border.  The  large  intestine  was  not 
congested,  the  contents  were  black;  the  small  intestine  was 
empty  and  flabby.  The  kidney,  pancreas  and  liver  were 
normal. 

In  both  dogs  the  stomach  and  intestinal  contents  and 
bile  were  saved  and  analyzed  for  the  presence  of  beryl- 
lium. In  the  first  animal  the  element  was  found  only  in  the 
feces  and  urine.  It  appeared  in  the  urine  voided  the  morn- 
ing after  the  operation  and  continued  to  be  excreted  during 
the  second  day  following.  In  the  second  experiment  traces 
of  beryllium  were  found  in  the  contents  of  both  the  large 
and  small  intestines  as  well  as  in  the  urine  and  feces.  It  ap- 
peared in  the  urine  voided  the  morning  following  the  oper- 
ation. The  volume  of  urine  excreted  the  day  following  the 
operation  was  largely  in  excess  of  the  normal  amount  for 
that  animal. 

Discussion  of  Rksults. — Practically  none  of  the  symptoms 
described  by  Siem  were  produced  in  these  experiments 
by  the  subcutaneous  injection  of  the  concentrated  solution. 


41 

The  animals  showed  a  temporary  soreness  in  the  area  of  in- 
jection and  stiffness  of  the  hind  leg  on  the  treated  side, 
but  this  soon  disappeared.  Very  marked  edema  and  ne- 
crosis set  in  within  two  or  three  days  after  the  injection  in 
all  the  animals  used  and  after  keeping  them  about  ten  days 
for  observ^ation  they  were  chloroformed.  The  beryllium 
sulfate  injected  in  the  concentrated  solutions  led  to  a  direct 
destructive  effect  upon  the  tissues  rather  than  a  systemic 
poisoning. 

It  was  found  that  the  Kahlbaum  preparation  of  beryllium 
sulfate  was  far  more  poisonous  in  its  effects  in  the  experi- 
ments with  ■intravenous  injections  than  the  tartrate  salt  used 
b}'  Siem.  In  the  second  experiment,  although  it  took  longer 
for  the  general  effects  to  be  produced,  they  were  more  severe 
in  the  end.  The  second  dog  was  apparently  recovering  from 
the  dose  the  day  before  the  final,  toxic  effects  were  the  most 
powerful. 

In  both  the  experiments  with  intravenous  injections 
the  symptoms  were  the  same.  The  beryllium  sulfate  had  a 
very  marked  effect  upon  the  heart,  making  the  action  very 
rapid,  irregular  and  weak.  During  the  operations  such 
extreme  muscular  trembling  occurred  that  before  the  animal 
was  removed  from  the  board  it  was  impossible  to  count  the 
pulse  or  respiration.  Directly  following  the  operation  there 
was  a  decided  increase  of  body  temperature  followed  by 
a  decrease  about  24  hours  before  the  death  of  the  animal. 
There  was  a  marked  increase  in  the  volume  of  urine.  In 
the  second  experiment  this  extra  elimination  amounted  to 
250  cc.  on  the  day  following  the  operation,  but  was  followed 
by  a  complete  retention. 

The  feces  were  bloody  and  diarrheal,  and  continued  so 
throughout  the  experiment.  There  were  no  evident  signs 
of  influence  on  the  senses  although  no  special  tests  were  made. 
In  neither  dog  were  there  convulsions. 

IV.  Effect  on  Enzyme  Action. 
I.  Effect  on  the  Activity  of  Sucrase. 
The  sucrase  was  precipitated  from  a  water  solution  of  yeast, 


42 

the  precipitate  washed  with  alcohol,  dissolved  in  distilled 
water  and  reprecipitated.  The  water  solution  of  the  second 
precipitate  was  used  in  the  experiment.  A  2  per  cent,  solution 
of  sucrose  free  from  reducing  substances  and  a  5  per  cent, 
solution  of  beryllium  sulfate  were  used.  The  tubes  were 
arranged  in  the  following  order. 

TABI.E   I. 


Control 

Control 

Control 

Tubes. 

I. 

II. 

III. 

IV. 

V. 

VI. 

VII. 

Sugar 

(2%) 

5  cc. 

5  cc. 

5  cc. 

5  cc. 

5  CC. 

5  cc. 

5  cc. 

Water 

3   " 

2    " 

I    " 

0   " 

0    " 

0  " 

4   " 

BeSO, 

(5%) 

I    " 

2   " 

3    " 

4   " 

5   " 

0  " 

0  " 

Enzyme 

solution 

I    " 

I   " 

I    " 

I    " 

0  " 

5   " 

I  " 

Per  cent. 

BeSO, 

0.5% 

10% 

1,5% 

2.0% 

0  " 

0  " 

0  " 

Results 

Heavy 

Good 

Slight  Very 

No  re- 

Heavy  Heavy 

reduc- 

reduc- 

reduc 

-  slight 

.  reduc- 

■    reduc- 

reduc- 

tion 

tion 

tion 

reduc 

-  tion 

tion 

tion 

tion 

The  tubes  were  placed  in  a  water  bath  at  38°  for  twenty 
minutes;  the  effect  of  the  beryllium  sulfate  was  tested  by 
comparing  the  relative  action  of  the  contents  of  the  different 
tubes  on  Benedict's  reducing  reagent.  The  results  showed 
that  there  was  a  slight  but  marked  decrease  in  reduction  as 
the  percentage  of  beryllium  sulfate  in  the  mixture  increased. 
An  amount  equal  to  1.5  per  cent,  in  the  solution  showed  a 
decided  inhibitory  effect. 

2.  Effect  on  the;  Activity  of  Ptyalin. 

To  a  I  per  cent,  solution  of  starch  paste  a  5  per  cent,  solution 
of  beryllium  sulfate  was  added  in  varying  proportions.  2  cc. 
of  filtered  saliva  were  added  to  each  tube  and  the  tubes  al- 
lowed to  stand  in  a  water  bath  at  38°  for  half  an  hour.  The 
effect  of  the  beryllium  sulfate  was  tested  by  comparing  the 
relative  power  of  the  solution  to  reduce  Benedict's  reagent. 
The  following  table  shows  the  proportion  of  the  mixtures  and 
the  results: 


43 


Table  II. 

Control 

Control        Control 

Tubes. 

I. 

11. 

III. 

IV. 

V. 

VI.               VII. 

Starch 

4  cc. 

4  cc. 

4.  cc 

4  cc. 

4  cc. 

4  CC.          4  CC. 

Water 

3    " 

2    " 

I    " 

0  " 

I    " 

1    "         5    " 

BeSO, 

(5%) 

I    " 

2    " 

3    " 

4   " 

5   " 

0  "        0   " 

Saliva 

2     " 

2    " 

2    " 

2    " 

0   " 

5   "         I    " 

Per  cent. 

BeSO, 

0.5% 

1.0% 

1.5% 

2,0% 

" 

0   "        0   " 

Results 

Slight 

Very 

No  re- 

No re- 

No re- 

Heavy Heavy 

reduc- 

slight 

duc- 

duc- 

duc- 

reduc- reduc- 

tion 

reduc- 
tion 

tion 

tion 

tion 

tion        tion 

The  inhibitory  effect  of  beryllium  sulfate  on  the  action  of 
saliva  was  very  marked.  The  tubes  were  allowed  to  stand 
for  two  hours  longer  but  practically  no  further  action  occurred. 
As  beryllium  sulfate  is  a  strongly  acid  salt  the  results  were 
as  would  be  expected  in  this  particular  case. 

3.  Effect  on  the  Activity  of  Pepsin. 

To  artificial  gastric  juice  (0.5  per  cent,  pepsin  in  0.2  per  cent, 
hydrochloric  acid)  the  5  per  cent,  solution  of  beryllium  sul- 
fate was  added  in  varying  proportions.  A  shred  of  fibrin 
was  added  to  each  tube  and  all  the  tubes  placed  in  a  water 
bath  at  38°  for  half  an  hour.  The  digestion  was  tested  by 
the  relative  disappearance  of  the  pieces  of  fibrin.  The  fol- 
f owing  table  gives  the  results: 


Tubes. 

Fibrin 
Water 
BeSO, 

(5%) 
Pepsin 
solution 
Per  cent. 

BeSO, 
Results 


I. 
Shred 
8  cc. 


0.5% 
Slight 
diges- 
tion 


II. 
Shred 
7  cc. 


Table  III. 

Control 
III.  IV.  V. 

Shred  Shred  Shred 
6  cc.     5  cc.     5  cc. 


Control 
VI. 

Shred 
5cc. 

o    " 
5   " 


10% 
No  di- 
ges- 
tion 


Control 
VII. 

Shred 
9  cc. 

o    " 

1  " 


1.5%  2.0%  o   "  o   "         o   " 

No  di-  No  di-  No  di-  Strong  Diges- 

ges-      ges-      ges-        diges-     tion 

tion      tion      tion        tion 


44 


At  the  end  of  half  an  hour  there  was  slight  digestion  in 
the  first  tube  and  in  the  control  but  practically  no  disinte- 
gration of  the  fibrin  in  any  of  the  other  tubes.  At  the  end 
of  two  hours  the  results  were  the  same.  This  experiment 
was  of  decided  interest  as  the  acid  reaction  of  the  beryllium 
sulfate  would  not  interfere  with  the  action  of  pepsin  as  it 
would  with  ptyalin. 

The  tubes  were  allowed  to  stand  for  24  hours  and  at  that 
time  digestion  had  occurred  in  all  the  tubes  containing  the 
pepsin  solution  and  beryllium  sulfate.  This  amount  of  di- 
gestion decreased  with  the  increase  percentage  of  beryllium 
sulfate.  There  evidently  was  a  marked  inhibitory  effect 
upon    the  enzyme  but  not  destruction. 

4.  Effect  on  the  Activity  of  Trypsin. 

Artificial  pancreatic  juice  was  made  by  extracting  the 
fresh  pancreas  with  30  per  cent,  alcohol.  The  extract  was 
carefully  neutralized  and  made  alkaline  with  0.5  per  cent. 
Na2C03.  The  5  per  cent,  solution  of  beryllium  sulfate  was 
added  in  varying  amounts  and  a  shred  of  fibrin  added  to 
each  tube.  All  tubes  were  allowed  to  stand  in  a  water  bath 
at  38°  for  thirty  minutes  and  the  digestion  determined  by 
the  relative  disintegration  of  the  fibrin. 

The  following  table  gives  the  conditions  and  results: 

Table  IV. 


Tubes. 

Fibrin 
Water 
BeSO, 

(5%) 
Trypsin 
solution 
Per  cent. 

BeSO^ 
Results 


Shred 
8  cc. 


Control 
11.            III.            IV             V. 

Control 
VI. 

Control 
VII. 

Shred  Shred  Shred  Shred 

Shred 

Shred 

7  cc.     6  cc.     5  cc.     5  cc. 

5  cc. 

9  cc. 

0.5% 

Very 
slight 
diges- 
tion 


1.0%  1.5%  2.0%  o   "  o   "        o   " 
No  di-  No  di-  No  di-  No  di-  Good     Slight 
ges-      ges-      ges-      ges-  diges-  diges- 
tion     tion      tion      tion  tion        tion 


At  the  end  of  half  an  hour  digestion  had  taken  place   in 
two  of  the  tubes  used  as   controls.     There  was  very  slight 


45 

digestion  in  the  first  tube  containing  the  beryllium  sulfate 
and  no  digestion  in  the  other  tubes.  After  allowing  the  tubes 
to  stand  24  hours  the  results  were  the  same. 

Conclusion. — It  is  evident  that  beryllium  sulfate  has  an 
inhibitory  effect  on  the  activity  of  all  the  enzymes  used  in 
the  above  experiments  but  sucrase  was  much  less  sensitive 
than  ptyalin,  pepsin  or  trypsin. 

V.  Precipitation  of  Proteins. 

The  effect  of  beryllium  sulfate  on  the  precipitation  of  pro- 
teins was  made  with  the  following  solutions: 

1.  Egg  Albumin. 

The  white  of  a  fresh  egg  was  diluted  with  an  equal  quantity 
of  distilled  water,  the  solution  passed  through  cheesecloth 
and  filtered.  A  5  per  cent,  solution  of  beryllium  sulfate  was 
added  a  drop  at  a  time.  Controls  were  made  with  5  per  cent, 
solution  of  sodium  chloride  and  ammonium  sulfate  and  i  per 
cent,  solution  of  picric  acid.  The  beryllium  sulfate  caused 
no  precipitation  of  the  albumin  but  compared  with  the  sodium 
.chloride  and  ammonium  sulfate  solutions  there  was  a  very 
slight  turbidity. 

2.  Mixed  Proteoses. 

Mixed  proteose  were  formed  by  peptic  digestion  of  meat. 
The  mixed  proteoses  were  precipitated  by  complete  saturation 
with  ammonium  sulfate  and  the  proteoses  dissolved  in  water. 
It  was  found  that  beryllium  sulfate  produced  no  precipitation 
of  these  products. 

3.  Gelatin. 

A  2  per  cent,  solution  of  gelatin  was  made  and  5  per  cent, 
solution  of  beryllium  sulfate  was  added,  i  cc.  at  a  time. 
There  was  no  precipitation  of  the  protein. 

4.  Acid  Meta-protein. 

The  blood  and  saline  material  were  removed  from  hashed 
lean  meat  by  allowing  the  meat  to  stand  in  running  water 
over  night.  At  the  end  of  that  time  the  meat  was  placed 
in  2  per  cent.  HCl  for  24  hours.  The  material  was  then 
squeezed  through  cheesecloth  and  filtered.  The  filtrate  con- 
taining  the   acid  meta-protein  was   brought  nearly    to   the 


46 

point  of  neutralization  and  the  solution  of  beryllium  sulfate 
added.  Controls  were  made  with  sodium  chloride  and  pic- 
ric acid.  The  beryllium  sulfate  produced  no  effect  upon  the 
precipitation  of  the  acid  meta-protein. 

Conclusion. — Proteins  are  not  precipitated  from  their 
solutions  by  beryllium  sulfate. 

VI.  Effect  on  Growth  of  Seedlings. 

Sistini^  performed  experiments  on  the  effect  of  beryllium 
on  the  growth  of  plants  by  replacing  the  magnesium  sulfate 
in  the  nutrient  solution  with  beryllium  sulfate.  The  solu- 
tions were  made  as  follows:  0.21  gram  potassium  chloride, 
0.46  gram  calcium  nitrate,  o.i  gram  iron  phosphate,  0.035 
gram  calcium  mono-phosphate  and  0.20  gram  of  magnesium 
sulfate  (or  0.18  gram  of  beryllium  sulfate).  The  solution 
was  made  up  to  a  liter.  This  gave  a  0.018  per  cent,  solution 
of  the  beryllium. 

He  states  in  his  results  that  beryllium  in  this  strength  may 
take  the  place  of  the  magnesium  in  the  growth  of  the  plant, 
but  is  not  a  complete  substitute  for  magnesium  in  the  pro- 
duction of  seeds.  The  plants  developed  and  bloomed  but 
the  seeds  were  not  fertile.  He  recovered  from  the  ash  of  the 
plant  2  per  cent,  of  the  beryllium  given  when  grown  in.  the 
nutrient  solution  and  traces  of  the  metal  in  the  ash  when 
the  plant  was  grown  in  earth  to  which  was  added  the  solution 
of  beryllium  sulfate. 

The  following  series  of  experiments  were  performed  to 
ascertain  the  effect  of  the  beryllium  sulfate  in  varying 
strengths  on  the  growth  of  the  seedlings.  Controls  were 
made  with  i  per  cent,  solution  of  sodium  chloride,  tap  water 
and  distilled  water. 

First  Experiment. — Lupine  seeds  were  soaked  in  water  for 
24  hours  and  grown  in  moss  for  48  hours.  At  the  end  of 
that  time,   15  mm.  was  measured    from  the  tip  of  the  root 

*  Sistini:  Ueber  einige  selten  in  Planzen  vorkoimnende  und  seither 
noch  nicht  darin  gefundene  chemische  Eleraente  Spiziell  iiber  Beryllium 
mit  Rucksicht  auf  einige  Kultivierte  Planzen.  Ckem.  Centr.,  1888,  p. 
1622.  Orig.  art.  Staz.  Spirem  Agrar,  xv,  p.  290-98  (19/10).  Pisa  lab  di. 
chimica  agraria.     Chem.  Centr.,  1888,  p.  1622. 


47 


and  that  distance  with  a  dot  of  India  ink.  The  seedlings 
were  then  suspended  in  the  different  solutions  in  such  a  man- 
ner that  the  root  was  immersed  exactly  to  the  distance  marked 
off.  The  growth  was  determined  by  the  difference  in  measure- 
ment from  the  ink  mark  to  the  tip  of  the  root.  The  differ- 
ence between  the  daily  measurement  and  the  original  15 
mm.  recorded  the  distance  of  growth.  Measurements  were 
taken  each  day.  In  the  following  experiment  the  dilution 
of  the  beryllium  sulfate  was  made  with  tap  water: 

Table  I. 


Tap 

Distil. 

NaCl 

BeSO* 

BeS04 

BeSO, 

water 

water 

1  % 

1  % 

0.8  % 

0.4  % 

growth. 

growth. 

growth. 

growth. 

growth. 

growth. 

Dates 

mm. 

mm. 

mm. 

mm. 

mm. 

mm. 

July 

31 

12  .0 

9  7 

1-5 

0.5 

I  .2 

2  .0 

Aug. 

15 

25   5 

21-5 

6.0 

I  .0 

I  .2 

2.0 

Aug. 

2 

33  6 

33-7 

6.4 

I  .2 

I    2 

2  .0 

Aug. 

15 

II  5 

9  5 

2  .0 

I  .0 

2-3 

2.0 

Aug. 

2 

23.2 

24.0 

6.2 

I  .0 

1-3 

15 

Aug. 

3 

40.6 

28.0 

6.2 

I  .0 

13 

1-5 

Aug. 

2 

12  .2 

9  7 

17 

0.2 

1 .0 

2.  2 

Aug. 

3 

23.2 

14.0 

6.5 

I  .0 

1 .0 

2.2 

Aug. 

4 

31,2 

33  7 

6.5 

1-5 

1 .0 

30 

Table  II. 

BeSO, 

BeSOi 

BeSOi 

BeS04 

BeSO* 

Tap 

sol. 

sol. 

•sol. 

sol. 

sol. 

Dates 

water. 

M/32. 

M/64. 

M/128. 

M/2S6. 

M/S12. 

Aug. 

6 

18.0 

I  .0 

I  .2 

15 

2-5 

6.7 

Aug. 

7 

42  .0 

I  .0 

15 

2-5 

4-7 

16.5 

Aug. 

8 

58.0 

I  .0 

30 

4-5 

4.2 

17  7 

Aug. 

7 

17.0 

I  .2 

I  .2 

2.2 

2  .0 

3-7 

Aug. 

8 

42  .0 

I  .0 

I  .  2 

30 

4.0 

15  2 

Aug. 

9 

53-7 

I  .0 

15 

3-2 

4-5 

18.7 

Aug. 

8 

18.0 

1.6 

2.6 

2.6 

2.0 

1,6 

Aug. 

9 

40.0 

I  .0 

2.6 

2.6 

2  .0 

1.6 

Aug. 

ID 

55  3 

(Shriv- 
elled) 
dead 

(Shriv- 
elled) 
dead 

2  .0 

2.0 

1.6 

Aug. 

9 

17  5 

1 .0 

15 

2.5 

2  .0 

2.0 

Aug. 

10 

42.0 

1 .0 

15 

2-3 

2  .0 

2.0 

Aug. 

II 

58  0 

(Shriv- 
elled) 

(Shriv- 
elled) 

2-3 

2.0 

2.0 

48 

Discussion  of  Results. — The  beryllium  sulfate  had  a  very- 
marked  effect  upon  the  growth  of  the  seedlings  as  shown 
by  the  tap  water  control.  In  many  cases  there  was  a  shrink- 
ing after  the  second  day.  In  every  case  in  which  the  beryl- 
lium sulfate  was  diluted  with  water  the  roots  blackened  and 
shrivelled  after  a  few  days.  In  the  stronger  solution  this 
effect  was  produced  within  24  hours. 

Second  Experiment. — The  following  experiments  were  made 
with  the  beryllium  sulfate  diluted  with  a  nutrient  solution^ 
instead  of  with  water.  The  nutrient  solution  was  made  as 
follows : 

6.0  grams  Ca(N03)2. 

1.5  grams  KNO3. 

1.5  grams  MgSO^. 

1.5  grams  neutral  potassium  phosphate. 

1.5  grams  sodium  chloride. 

600  cc.  distilled  water. 


Table  III. 

Date. 

Control 
nut.  sol. 

BeSOi      BeSOi 
sol.           sol. 

M/2.          M/4. 

BeS04 
sol. 

M/8. 

BeS04 

sol. 
M/16. 

BeS04 
sol. 

M/32. 

Aug. 

II 

II. 2 

0.0          1.7 

2.0 

15 

2.0 

Aug. 

12 

48.0 

I  .0          2.0 

1-7 

15 

2.0 

Aug. 

13 

64.0 

I .0          2.0 

1-7 

1-5 

2.0 

Aug. 

13 

14.0 

0.5         I.O 

1-3 

1-5 

2.0 

Aug. 

14 

530 

I  .0          1.0 

15 

1-7 

2.0 

Aug. 

15 

64.0 

1.0          1.0 

1-3 

17 

2.0 

In  the  two  experiments  in  which  nutrient  solution  was 
used  instead  of  tap  water  the  roots  remained  white  and  crisp 
for  several  days  although  the  beryllium  sulfate  entirely  in- 
hibited growth. 

Conclusion.  —  From  the  experiments  carried  on  in  the 
manner  described  above  beryllium  sulfate  to  the  strength 
of  a  M/512  solution  had  a  marked  inhibiting  effect  on  the 
growth  of  lupin  seeds. 

Third  Experiment. — Timothy  seed  was  allowed  to  sprout 
^  McDougal:  "Practical  Text-book  of  Plant  Physiology,"  1911,  p.  224. 


49 


on  several  pieces  of  wet  blotting  paper.  When  the  growth 
had  reached  the  height  of  i/8  inch,  lo  drops  of  the  following 
strengths  of  beryllium  sulfate  were  added  each  day  and  at 
the  end  of  a  week  the  heights  of  the  growths  on  the  various 
pieces  of  blotting  paper  compared. 


Strength  of 
BeSO^. 

Results 


Table  IV. 


Control 
tap  water. 


BeSO* 

M/256. 


BeSO* 

M/512. 


BeS04 
M/1024. 


BeS04 
M/2048. 


BeSOi 
M/4096. 


Good      Slight  Growth   Good     Good     Good 
growth  growth  growth  growth  growth 


Table  V, 

strength 
of  BeS04. 

Control 
tap  water. 

BeSO*.          BeSO,. 
Per  cent.      Per  cent. 

BeSO*. 
Per  cent. 

BeSO*. 
Per  cent. 

BeSO«. 

Results 

Good 
growth 

0.5            0.25 

Slight     Growth 
growth 

0.125 

Good 
growth 

0.06 

Good 
growth 

0.03 
Good 
growth 

Discussion  of  Results. — Comparing  the  growth  of  the 
grass  for  the  various  strengths  of  the  beryllium  sulfate  solu- 
tions with  the  tap  water  control  the  M/256  solution  showed 
a  very  decided  inhibitory  effect  and  the  M/512  solution  a 
detrimental  effect.  Beyond  this  dilution  the  beryllium  sul- 
fate had  little  inhibitory  effect  on  the  growth. 

It  was  very  noticeable  that  when  the  experiment  was 
finished  and  the  plots  of  grass  allowed  to  remain  without 
attention,  the  plants  that  had  been  treated  with  beryllium 
sulfate  became  withered  and  yellow  within  24  hours  while 
the  control  which  had  received  only  tap  water  remained 
green  and  fresh  for  more  than  48  hours. 

VII.     Effect  on  Bacterial  Growth. 

The  bactericidal  property  of  beryllium  sulfate  was  tested 
by  its  effect  on  the  growth  of  Staphylococcus  pyogenes  and  B. 
coli  communis.  Tubes  containing  10  cc.  of  gelatin  medium 
were  inoculated  by  adding  i  cc.  of  an  emulsion  of  the  bacteria. 
I  %  solution  of  BeSO^  was  added  in  varying  amounts  to  dif- 
ferent tubes  and  the  tubes  plated.     Colonies  developed  on  all 


50 

the  plates  up  to  the  one  to  which  lo  cc.  of  the  BeSO^  had  been 
added.  As  the  beryllium  sulfate  is  strongly  acid  in  its  reaction 
the  inhibitory  effect  produced  at  this  point  may  have  been  due 
to  the  acidity. 

VIII.     Summary  of  Conclusions. 

The  experiments  with  beryllium  sulfate  have  shown  very 
conclusively  that  the  substance  has  a  marked  effect  on  bio- 
chemical processes.  When  administered  with  the  food  it 
produced  decided  nutritive  disturbance,  which  manifested 
itself  in  loss  of  body  weight,  total  inorganic  matter,  nitrogen, 
sulfur  and  phosphorus. 

When  large  doses  were  administered  per  os  the  substance 
caused  vomiting  in  the  dog  before  a  sufficient  amount  was 
absorbed  to  produce  any  other  obvious  toxic  symptoms. 

When  the  calculated  lethal  dose  was  administered  by  a 
single  subcutaneous  injection,  the  substance  produced  edema 
and  necrosis  of  the  tissue  extending  over  a  large  area.  No 
other  decided  symptoms  were  produced  by  this  method. 

Very  gradual  intravenous  injections  of  the  salt  produced 
decided  toxic  effect.  The  action  of  the  heart  became  irregular 
— usually  rapid  and  very  weak;  the  respiration  also  became 
irregular  and  shallow.  During  the  period  of  injection  it  pro- 
duced decided  muscular  trembling  but  this  disappeared  soon 
after  the  operation.  As  a  direct  effect  of  the  injection  the 
temperature  increased,  sometimes  to  105°,  but  about  24  hours 
before  the  death  of  the  animal  the  temperature  began  to 
decrease  and  steadily  fell. 

After  the  intravenous  injection  there  was  an  increased  elimi- 
nation of  urine  followed  by  retention.  The  feces  became 
diarrheal  and  bloody.  Vomiting  began  about  the  time  the  dog 
refused  food  or  water. 

The  beryllium  sulfate  had  a  decided  inhibitory  effect  on  the 
action  of  ptyalin,  pepsin,  and  trypsin.  It  also  retarded  the 
action  of  sucrase  but  not  to  so  great  an  extent.  Solutions  of 
the  salt  ( I  %  or  less)  did  not  effect  the  precipitation  of  proteins 
from  neutral  or  acid  solutions. 


51 

Below  the  concentration  of  M/512  solution,  the  beryllium 
sulfate  did  not  have  an  inhibiting  action  on  the  growth  of 
lupine  or  timothy  seedlings,  but  the  more  concentrated  solutions 
of  this  salt  prevented  growth  and  had  a  destructive  effect  on 
the  plant  life. 

Berylhtun  sulfate  when  present  in  proportions  less  than  0.5% 
had  very  little,  if  any,  bactericidal  action. 


BIOGRAPHICAL. 

Emily  Cromwell  Seaman  was  bom  in  Brooklyn,  New  York, 
on  March  5th,  1876.  She  received  the  degree  of  B.S.  from 
Adelphi  College  in  1899,  and  the  degree  of  M.  A.  from  Columbia 
University  in  1905.  From  1901  to  1908  she  taught  in  elemen- 
tary schools  in  Brooklyn.  She  resigned  this  work  to  specialize 
in  Physiological  Chemistry  and  entered  the  Laboratory  of  Bio- 
logical Chemistry  of  Columbia  University  in  the  Spring  of  1908. 
In  1909  she  was  given  the  position  of  Tutor  in  Physiological 
Chemistry  at  Teachers  College  and  in  1910  was  appointed  In- 
structor. 


\/ 


BIBLIOGRAPHY. 

Blake:  Ueber  den  Zusammenhang  der  Molekularen  Eigenschaften  anorgan- 
ischer  Verbindungen  und  ihre  Wirkung  auf  den  lebenden  Thierischen 
Organismus.     Berichte  der  deut.  chem.  Gessellschaft,  xiv,  p.  394. 

Blake:  Atomic  Weight  of  Beryllium  as  Determined  by  its  Physiological 
Actions.     Chemical  News,  Ixv,  p.  iii. 

Brunton  and  Cash:  Royal  Society,  Philosophical  Transactions,  1884,  p. 
197. 

Siem:  Inaug.  Dissert.,  Dorpat,  1886.  Ueber  die  Wirkung  des  Aluminium 
und  des  Berylliums  auf  den  thierischen  Organismus. 

Sistini:  Orig.  art.  Staz.  Spirem  Agrar,  xv,  p.  290-298  [19/10].  Pisa  lab. 
di  chimica  agraria.  Chem.  Centr.,  1888,  p.  1622.  Ueber  einige  selten 
in  Planzen  vorkommende  und  seither  noch  nicht  darin  gefiuidene 
Chemische  Elemente  Spiziell  iiber  Beryllium  mit  Rucksicht  auf  einige 
Kultivierte  Planzen. 


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